生物活性
Dabrafenib (GSK2118436)是一种突变型BRAFV600特异性抑制剂,IC50为0.8 nM,作用于B-Raf(wt)和c-Raf效果低4和6倍。Dabrafenib对Raf激酶具有选择性,对B-Raf的活性比其它测试过的91%的激酶高400倍。 Dabrafenib抑制B-RafV600E激酶,导致ERK磷酸化降低和抑制细胞的增值,在特异性编码突变的B-RafV600E的癌细胞中细胞停滞在G1期。
使用AbMole产品发表的文献
产品使用成果展示
|
数据来源 |
Int J Mol Sci (2018). Figure 5. GSK2118436A (AbMole BioScience, Houston, TX, USA) |
方法 |
vitro experiments |
细胞系/动物模型 |
CAR-T cells |
浓度 |
1 μM |
处理时间 |
16 h |
实验结果 |
CAR-T cells in the conditions without inhibitor, with DMSO solvent control, with Vem alone, Tram alone, Dabra alone, and the combination Dabra + Tram |
|
数据来源 |
Int J Mol Sci (2018). Figure 4. GSK2118436A (AbMole BioScience, Houston, TX, USA) |
方法 |
vitro experiments |
细胞系/动物模型 |
CAR-T cells |
浓度 |
1 μM |
处理时间 |
16 h |
实验结果 |
The presence of Vem alone, Tram alone,Cobi alone, Vem + Cobi, and Dabra + Tram, but not of Dabra alone seemed to reduce these quantities to approximately 50%. |
|
数据来源 |
Int J Mol Sci (2018). Figure 3. GSK2118436A (AbMole BioScience, Houston, TX, USA) |
方法 |
vitro experiments |
细胞系/动物模型 |
CAR-T cells |
浓度 |
1 μM |
处理时间 |
16 h |
实验结果 |
"The condition with Vem + Cobi was similarly inhibited as Vem alone, while the Dabra + Tram condition
was significantly less inhibited" |
|
数据来源 |
Int J Mol Sci (2018). Figure 2. GSK2118436A (AbMole BioScience, Houston, TX, USA) |
方法 |
vitro experiments |
细胞系/动物模型 |
CAR-T cells |
浓度 |
1 μM |
处理时间 |
16 h |
实验结果 |
Dabra alone had a significantly weaker effect on CD25-upregulation than Vem alone, and the combination Dabra + Tram resulted in the mildest effects on CD25 upregulation |
|
数据来源 |
Journal of Cell Science (2016) . Figure 3. Dabrafenib (M1988, Abmole, Houston, TX) |
方法 |
Cell apoptosis assay and western blot |
细胞系/动物模型 |
IM9 and RPMI8226 cells expressing PAX5 |
浓度 |
10 μM |
处理时间 |
30 h |
实验结果 |
Similarly, the selective RIP2 inhibitor SB 203580, but not the RIP1 inhibitor Necrostatin-1 (Degterev et al., 2013) nor the RIP3 inhibitor Dabrafenib (Li et al., 2014), was able to significantly increase Bortezomib-induced apoptosis in IM9 andPAX5-expressing RPMI8226 cells (Fig. 3D). Furthermore, only SB 203580 decreased Bortezomib-induced activation of RIP2 and NF-κB in IM9 cells (Fig. 3F). |
|
数据来源 |
Cell Report (2016). Figure 7.dabrafenib (GSK2118436, Abmole) |
方法 |
Tumour volume and Immunoblotting |
细胞系/动物模型 |
|
浓度 |
30 mg/kg |
处理时间 |
25~30 days |
实验结果 |
BRAFV600E/DK is Responsible for Resistance to BRAFi.Both PDXs were treated either with the BRAF inhibitor dabrafenib, to which they were completely resistant (Figures 7G and 7H). Indeed, AKT signaling was much more active in the M048R2.X2 PDX (Figure 7I), explaining its only partial sensitivity to LY3009120. |
|
数据来源 |
Cell Report (2016). Figure 3.dabrafenib (GSK2118436, Abmole) |
方法 |
Immunoblotting and qPCR |
细胞系/动物模型 |
|
浓度 |
30 mg/kg |
处理时间 |
25~30 days |
实验结果 |
These results demonstrate concordance between drug responses in patients and their corresponding PDXs, and they illustrate that the therapy response can be either stable or dynamic. |
|
数据来源 |
Cell Report (2014) . Figure 7.Dabrafenib (Abmole) |
方法 |
In Vivo Experiments |
细胞系/动物模型 |
M026R.X1 tumors |
浓度 |
30mg/kg |
处理时间 |
33 days |
实验结果 |
Synthetic lethality by hypoxia induction and Chek1/2 inhibition in vivo was observed for the PDX derived from a melanoma patient who had acquired resistance to BRAF inhibition. |
|
数据来源 |
EMBO Molecular Medicine (2015) . Figure 4.Dabrafenib (GSK2118436, Abmole) |
方法 |
Mice--Derived Xenografts |
细胞系/动物模型 |
A375 cells infected with pQXCIPGFP, MEK1WT, and MEK1T55delinsRT |
浓度 |
30 mg/kg daily by oral gavage |
处理时间 |
12 days |
实验结果 |
Altogether, these data confirm that MEK1T55delinsRT confers resistance to BRAF inhibition in vivo and induces local invasion. |
|
数据来源 |
EMBO Molecular Medicine (2015) . Figure 4.Dabrafenib (GSK2118436, Abmole) |
方法 |
CellTiter-Blue Cell Viability Assay (G8081, Promega) |
细胞系/动物模型 |
A375-GFP, 888Mel-MEK1T55delinsRT,SK-MEL-28 |
浓度 |
0, 0.25, 0.5, 1, 2.5, 5 µ M |
处理时间 |
5 days |
实验结果 |
MEK1T55delinsRT-expressing cells were also more resistant to low levels of the ERK inhibitor SCH772984 than control populations (Fig 4F and G). |
实验参考
体外实验* |
细胞系 |
A375-GFP, 888Mel-MEK1T55delinsRT,SK-MEL-28 |
方法 |
Dose–response curves were performed as follows: 3,000–6,000 cells were plated per well in a 96-well plate. For each condition, triplicates were plated. The next day, drug was added to the wells. At day 5 of the assay, medium-containing drug was removed and survival was measured by CellTiter-Blue Cell Viability Assay (G8081, Promega).
|
浓度 |
0, 0.25, 0.5, 1, 2.5, 5 µ M |
处理时间 |
5 Days |
*上述方法来自公开文献,仅供相同目的实验参考。如实验目的、材料、方法不同,请参考其他文献。
体内实验* |
动物模型 |
A375 cells infected with pQXCIPGFP, MEK1WT, and MEK1T55delinsRT Mice--Derived Xenografts |
配制 |
Dabrafenib powder was first dissolved in DMSO and consequently, before injection, dissolved in 0.5% hydroxypropylmethylcellulose (Sigma-Aldrich), 0.2% Tween-80 in pH 8.0 distilled H2O. |
剂量 |
30 mg/kg daily (6 days per week) |
给药处理 |
oral gavage |
*上述方法来自公开文献,仅供相同目的实验参考。如实验目的、材料、方法不同,请参考其他文献。
化学性质
分子量 |
519.56 |
分子式 |
C23H20F3N5O2S2 |
CAS号 |
1195765-45-7
|
溶解性(25°C) |
DMSO 30 mg/mL |
储存条件 |
粉末型式 -20°C 3年;4°C 2年
溶于溶剂 -80°C 6个月;-20°C 1个月
|
运输方式 |
冰袋运输,根据产品的不同,可能会有相应调整。 |
储备液配制
*下述溶液配置方法仅为基于分子量计算出的理论值。不同产品在配置溶液前,需考虑其在不同溶剂中的溶解度限制。
Concentration / Solvent Volume / Mass |
1 mg |
5 mg |
10 mg |
1 mM |
1.9247 mL |
9.6235 mL |
19.2471 mL |
5 mM |
0.3849 mL |
1.9247 mL |
3.8494 mL |
10 mM |
0.1925 mL |
0.9624 mL |
1.9247 mL |
不同实验动物依据体表面积的等效剂量转换表(参考来源于公开文献)
|
小鼠 |
大鼠 |
兔 |
豚鼠 |
仓鼠 |
狗 |
重量 (kg) |
0.02 |
0.15 |
1.8 |
0.4 |
0.08 |
10 |
体表面积 (m2) |
0.007 |
0.025 |
0.15 |
0.05 |
0.02 |
0.5 |
Km 系数 |
3 |
6 |
12 |
8 |
5 |
20 |
动物 A (mg/kg) = 动物 B (mg/kg) × |
动物 B的Km系数
|
动物 A的Km系数 |
例如,依据体表面积折算法,将化合物用于小鼠的剂量20 mg/kg 换算成大鼠的剂量,需要将20 mg/kg 乘以小鼠的Km系数(3),再除以大鼠的Km系数(6),得到化合物用于大鼠的等效剂量为10 mg/kg。
参考文献
[1] Greger JG, et al. Mol Cancer Ther. Combinations of BRAF, MEK, and PI3K/mTOR inhibitors overcome acquired resistance to the BRAF inhibitor GSK2118436 dabrafenib, mediated by NRAS or MEK mutations.
[2] Hong DS, et al. Clin Cancer Res. BRAF(V600) inhibitor GSK2118436 targeted inhibition of mutant BRAF in cancer patients does not impair overall immune competency.